Method for determining likelihood of an inflammatory gastrointestinal tract disease

Abstract

The aspects of the disclosed embodiments relate to method for determining a likelihood of an inflammatory gastrointestinal tract disease. The method includes diluting a biological sample of a human subject and contacting it with 8-anilinonaphthalene-1-sulfonic acid as modulating agent and further with a reagent, said reagent comprising a peroxidase enzyme and a label selected from a europium chloride and terbium chloride. The sample is incubated and excited, and the time-resolved luminescence signal of the label in the sample is measured. If the luminescence signal is at least 112% higher than for a control sample from a human so subject without of an inflammatory intestine disease, an increased likelihood of an inflammatory intestine disease of the human subject is determined.

The invention claimed is: 
     
       1. A method for determining likelihood of an inflammatory gastrointestinal tract disease, comprising
 diluting a biological sample of a human subject in an aqueous solution to obtain a diluted sample; 
 contacting the diluted sample with 8-anilinonaphthalene-1-sulfonic acid as modulating agent to obtain a pre-measurement sample; 
 forming a measurement sample by contacting the pre-measurement sample with a reagent, said reagent comprising a peroxidase enzyme and a label selected from europium chloride and terbium chloride; 
 incubating the measurement sample; 
 exciting the measurement sample; and 
 measuring time-resolved luminescence signal of the label in the measurement sample and determining an increased likelihood of an inflammatory intestine disease of the human subject if the luminescence signal is at least 112 percent (-%) higher than for a control sample from a human subject without an inflammatory intestine disease. 
 
     
     
       2. The method according to  claim 1 , wherein the time-resolved luminescence signal is measured for a time of 200-8000 μs after a 200-8000 μs delay time. 
     
     
       3. The method according to  claim 1 , wherein the peroxidase enzyme is selected from a group consisting of horse radish peroxidase, ascorbate peroxidase, chloride peroxidase, cytochrome c peroxidase, haloperoxidase, lactoperoxidase, myeloperoxidase and mixtures thereof. 
     
     
       4. The method according to  claim 1 , wherein a solvent is added to the measurement sample before incubation, the solvent being selected from dimethyl sulfoxide, N,N-dimethylformamide, diisopropyl ether, and mixtures thereof. 
     
     
       5. The method according to  claim 1 , wherein the label is used in an amount of 4 μL of label mixture containing europium chloride 0.717 μM, trioctylphosphine oxide 0.430 μM and nitrilotriacetic acid 0.430 μM (NTA) in dimethyl sulfoxide, and the peroxidase enzyme is used in an amount of 4 μl of 2.5 units per mg diluted in 1 mL of physiological salt solution. 
     
     
       6. The method according to  claim 1 , wherein the label is europium chloride label containing europium chloride, nitrilotriacetic acid and trioctylphosphine oxide. 
     
     
       7. The method according to  claim 1 , wherein incubation time is 5-20 minutes. 
     
     
       8. The method according to  claim 1 , wherein 8-anilinonaphthalene-1-sulfonic acid is used in an amount of 4 μL of a solution having a concentration of 10 μM-2000 μM. 
     
     
       9. The method according to  claim 1 , wherein the biological sample is diluted to a 5-1000 fold dilution. 
     
     
       10. The method according to  claim 1 , wherein the biological sample is a faecal sample, saliva sample, urine sample, blood sample or a bile sample. 
     
     
       11. The method of  claim 1 , wherein contacting the diluted sample with 8-anilinonaphthalene-1-sulfonic acid is performed ex vivo.